PMID- 34859108 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220428 IS - 2321-3868 (Print) IS - 2321-3876 (Electronic) IS - 2321-3868 (Linking) VI - 9 DP - 2021 TI - Overexpression of miR-101 suppresses collagen synthesis by targeting EZH2 in hypertrophic scar fibroblasts. PG - tkab038 LID - 10.1093/burnst/tkab038 [doi] LID - tkab038 AB - BACKGROUND: MicroRNA-101 (miR-101) is a tumor suppressor microRNA (miRNA) and its loss is associated with the occurrence and progression of various diseases. However, the biological function and target of miR-101 in the pathogenesis of hypertrophic scars (HS) remains unknown. METHODS: We harvested HS and paired normal skin (NS) tissue samples from patients and cultured their fibroblasts (HSF and NSF, respectively). We used quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), fluorescence in situ hybridization (FISH), enzyme-linked immunosorbent assays (ELISA) and Western blot analyses to measure mRNA levels and protein expression of miR-101, enhancer of zeste homolog 2 (EZH2), collagen 1 and 3 (Col1 and Col3) and alpha-smooth muscle actin (alpha-SMA) in different in vitro conditions. We also used RNA sequencing to evaluate the relevant signaling pathways and bioinformatics analysis and dual-luciferase reporter assays to predict miR-101 targets. We utilized a bleomycin-induced fibrosis mouse model in which we injected miR-101 mimics to evaluate collagen deposition in vivo. RESULTS: We found low expression of miR-101 in HS and HSF compared to NS and NSF. Overexpressing miR-101 decreased Col1, Col3 and alpha-SMA expression in HSF. We detected high expression of EZH2 in HS and HSF. Knockdown of EZH2 decreased Col1, Col3 and alpha-SMA in HSF. Mechanistically, miR-101 targeted the 3'-untranslated region (3'UTR) of EZH2, as indicated by the decreased expression of EZH2. Overexpressing EZH2 rescued miR-101-induced collagen repression. MiR-101 mimics effectively suppressed collagen deposition in the bleomycin-induced fibrosis mouse model. CONCLUSIONS: Our data reveal that miR-101 targets EZH2 in HS collagen production, providing new insight into the pathological mechanisms underlying HS formation. CI - (c) The Author(s) 2021. Published by Oxford University Press. FAU - Li, Jie AU - Li J AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Li, Yan AU - Li Y AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Wang, Yunchuan AU - Wang Y AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - He, Xiang AU - He X AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Wang, Jing AU - Wang J AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Cai, Weixia AU - Cai W AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Jia, Yanhui AU - Jia Y AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Xiao, Dan AU - Xiao D AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Zhang, Jian AU - Zhang J AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Zhao, Ming AU - Zhao M AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Shen, Kuo AU - Shen K AUID- ORCID: 0000-0001-8803-6507 AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Li, Zichao AU - Li Z AUID- ORCID: 0000-0002-4679-1042 AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Jia, Wenbin AU - Jia W AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Wang, Kejia AU - Wang K AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Zhang, Yue AU - Zhang Y AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Su, Linlin AU - Su L AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Zhu, Huayu AU - Zhu H AUID- ORCID: 0000-0002-3038-6624 AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. FAU - Hu, Dahai AU - Hu D AD - Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China. LA - eng PT - Journal Article DEP - 20211126 PL - England TA - Burns Trauma JT - Burns & trauma JID - 101651457 PMC - PMC8633590 OTO - NOTNLM OT - Collagen OT - EZH2 OT - Fibroblasts OT - Hypertrophic scars OT - Skin OT - miR-101 EDAT- 2021/12/04 06:00 MHDA- 2021/12/04 06:01 PMCR- 2021/11/26 CRDT- 2021/12/03 07:09 PHST- 2021/05/23 00:00 [received] PHST- 2021/07/22 00:00 [revised] PHST- 2021/12/03 07:09 [entrez] PHST- 2021/12/04 06:00 [pubmed] PHST- 2021/12/04 06:01 [medline] PHST- 2021/11/26 00:00 [pmc-release] AID - tkab038 [pii] AID - 10.1093/burnst/tkab038 [doi] PST - epublish SO - Burns Trauma. 2021 Nov 26;9:tkab038. doi: 10.1093/burnst/tkab038. eCollection 2021.